Download evaluation the immune status of the burn patients infected with

International Journal of Research and Development in Pharmacy and Life Sciences
Available online at http//www.ijrdpl.com
June - July, 2016, Vol. 5, No.4, pp 2219-2225
ISSN (P): 2393-932X, ISSN (E): 2278-0238
Research Article
EVALUATION THE IMMUNE STATUS OF THE BURN PATIENTS INFECTED WITH BACTERIA
PSEUDOMONAS AERUGINOSA IN KARBALA CITY
Dhawaa mohammed alkateeb1*, Wafaa sadiq alwazni1, Yasemin khudair alghanimi2, Muneer abdrasul altuoma3
1.
Karbala University, Faculty of Science, Department of Life Sciences, Iraq
2.
Karbala University, Faculty of Education, Department of Life Sciences Hussein, Iraq
3.
Ministry of Health, laboratory Hussein Teaching Hospital, Iraq
*Corresponding author’s Email: [email protected]
(Received: April 17, 2016; Accepted: May 20, 2016)
ABSTRACT
The present study aimed to release frequency pseudomonas aeruginosa bacterium Pseudomonas aeruginosa among burns patients in the province of
Karbala ratio. With shown immunological changes to the patient due to various injuries, burns with the aggravation of the injury that the germ and the evolution of
the case of septicemia and death, so it was in the period from 08/16/2014 to -16 2-2014 Leather collect swabs from 64 patients of burn patients admitted to the
Hussein Teaching Hospital In Karbala, and after that the cultivation of P.aeruginosa isolated by 45% of all burns swabs Group. When examining some
immunological criteria for patients with these bacteria Was clearly evident and clear rise in the number of white blood cells in patients where proportional rise with
the degree of burning and intensity, reaching the preparation of cells to mm3) / 103) 10.44, 13.222 and 15.955 for each of the patients with simple and
moderate to severe burns group, respectively, as was the rise in particular for neutrophil white blood cells compared to other cellular species.
The concentrations of both acute phase protein and supplemented significant increase was found in the acute phase Brocaat values and which is growing
directly proportional to the severity of the burn for up to 63.4 in simple burns while reached 95.2 and 121.8 for each of moderate to severe burns respectively.
The concentrations of complement protein C3 has statistical analyzes showed a non-significant increase complement each level of the third degree burns and
Statistics by mgldL 125.45, medium and severe 137.5 mgldL 140.15 mgldL compared with healthy people 122.32 mgldL. While was significant rise in motor values
cellular IL-6 for up to 118 pg / ml, 86 pg / ml, 33 pg / ml for each of the patients Statistics burns and moderate to severe respectively. On the contrary, it has
recorded kinetic values cellular TNF higher higher in patients with severe burns, reaching 600.45 pg / ml compared to people healthy 175.15 pg / ml..
Keywords: Pseudomonas aeruginosa, burn patients, Brocaat values, complement protein C3.
INTRODUCTION
changes the main reasons that make the patient susceptible
It provided burns and is one of the most forms of skin bruise
to blood sepsis where you get an increase in the
time period as a result of the loss of his job and the defense,
effectiveness of the macrophage after obtaining thermal
which result in the possibility of injury septic therefore require
injury severe, leading to increase the effectiveness of those
rapid intervention needed to reduce injuries and deaths
cells, which in turn lead to stimulate the inflammatory
among burn patients for medical care (Ekrami & kalantar,
mediators initial production (Proinflammatory mediators) such
2007). A patient who suffered a severe thermal injury burns,
as prostaglandins, TNF, IL-6, IL-1 and other inflammatory
especially second-degree burns and the third suffers to large
factors other also to increase these factors role in
immunological and physiological changes where it is immune
suppressing the immune system cells and thus the patient
©SRDE Group, All Rights Reserved.
Int. J. Res. Dev. Pharm. L. Sci.
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Alkateeb D. M. et al., June - July, 2016, 5(4), 2219-2225
becomes susceptible to injury bloody sepsis as a result of
one of the immune proteins as a level less than 1 mg / L in
what is accompanied by the burning of failure in many
the normal case and its level rises in the acute inflammatory
events physiological members of the patient's body, which is
conditions is evidence of inflammation (Yu, 2014).
considered one of the major causes of morbidity and death
Adding that the level of the complement proteins significantly
in these patients (Meakins, 1990 Harris & Gelfand, 1995;
change
Yamada etal., 2000;).
proportion to the severity of the burn and then return to rise
Upon entering the strange nurse to the human body, the
above the natural level of it after a limited period as the
immune system to distinguish those nurses through receptors
increase C5a levels and C3a result in changes in blood
on the surface of macrophages of cells granular A
pressure and vascular permeability as well as significant
granulocyte cells only Monocyte cells, Macrophage, which is
changes in the the functions of the egg cells, it was found that
the first step to create an immune response against the nurses
small amounts of the complement protein C5a leads to
cells, while the second step is to activate all of the tracks
increase the functional activity of the cells of eggs, while the
Complement
Complement
system))
inflammatory
excess of it amounts lead to curbed and this thing also
mediators Interlukin)) that operate on the overlap and
applies to the C3 b (Bengtson and Heideman, 1987; Yurt
organize the work of immune cells specialized, such as white
and Pruitt, 1986).
blood cells neutrophil cells and macrophage then activate
Play other immune proteins known as interleukins an
the immune response of cellular gained and immune cell
important role in determining the effectiveness of the immune
helper T first two types, and the second (T helper cell 1 & 2)
system in patients with burns like interleukin 6 (IL-6), which
(Sadikot et al; 2005).
give rise for glycoprotein Glycoprotein molecular weight of
In thermal injuries, burns contaminated with germs note there
26 kDa and is produced from a single cell nucleus,
is an increase in the number of white blood cells
macrophage , lymphocytes , endothelial cells , cells of B and
(Leukocytosis) to attack the bacteria involved in the thing's
T and the cornea cell in skin and others. Future IL-6R consists
body infected, which depends on the presence of high
of two series of proteins first unit low familiarity series Alpha
movement of these defensive cells and they are attracted to
(chain) output and the complex will be linked with beta chain
the site of the injury quickly, leading to a decrease in these
(chain) in excess of the value of linking and transmission of
blood cells in the bloodstream thing which makes the body
signals into the cell When for damage in the tissues of the
has difficulty in getting rid of pathogenic microbes hyphen to
body injury will begin of Cellular immunity by inflammation
the blood thus the deterioration of the patient's condition and
of topical response through the ability to Langerhans island
his death( Gamelli et al., 1986)).
cells in cooperation with the T and its neighboring
acute phase protein one of the proteins that increase their
keratinocytes in the epidermis area norepinephrine many
level during the presence of inflammation in the body, so the
immune components such as IL-6, which has an important role
high level of this protein in Patients serum is a sign of the
in the body's immunity and to be those components immune to
presence of inflammation as the acute phase proteins are a
the skin, called tissue fibro-related skin device (Parslow et
group of proteins that already exist in the plasma of human
al., 2001). The Alpha TNF is a mediator inflammatory first
blood low concentrations susceptibility to sedimentation the
Proinflammatory liberated mainly of the only machrophage
presence of calcium and increasing emancipation under
cells that are the main source of liberation. There is also a
certain conditions such as tumors and tissue damage burns
wide range of cells can heading such as mast cells and cells
and injuries germ even respiratory effects possible affect on
lymphatic T, B, NK cells NK, cells and neutrophil and
levels in human blood plasma (Dannacco & sansonno, 2005).
endothelial cells and smooth muscle and cardiac fibroblasts
CRP and acute phase proteins and the detachment of the
and bone-building cells and this factor as one of the
cells of the liver inflammation due to infection, which affects
important media is to transmit signals and perform many of
the body causing raise the levels of cytokines produced by
the immunological events that outgoing signals result in a
macrophages in the blood. In addition to that this protein is
wide range of cellular responses of programmed cell
©SRDE Group, All Rights Reserved.
and
after various burns where reduced its focus in
Int. J. Res. Dev. Pharm. L. Sci.
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Alkateeb D. M. et al., June - July, 2016, 5(4), 2219-2225
death(Bouwmeester et al., 2003; Brodley & combridge). In
biochemically confirmed the results of these tests using the
general cause immune to decline due to severe burns
system 20 and Api Alvaateix
fundamental changes in the balance between the helper cells
Calculation of the total count and differential white blood
ratio of the first to the second helper cells occur as well as
cells
between the percentage of cells lymph assistance (CD4 TH -
Total count of WBC
lymphocytes) cells ballasts (CD8 suppresser T cell) (Guo et al
Use the full complet blood count device (CBC) used in the
0.2003). Therefore During the first week of burning
blood disease unit at Al-Hussein Teaching Hospital for the
observed decline in the totals Bcell B lymphocytes that are
analysis of blood and automatically calculate the total count
commensurate with the severity of the burn, leading to
and differential of white blood cells
reduced production immune IgG that does not get low in the
Identify some of the humoral immune parameters such as:
formation of antibodies and other B-cells by itself and in
• C-reactive protein of Boditech company to measure the
these
qualitative part of the effective protein.
circumstances
be
the
body's
reaction
firing
(Prostoglandin) coupled with the low level of IL-12 which has
• measuring the serum level of the complement proteins C3
a synergistic effect in the differentiation of helper cells first
and by spreading the immune radiographic single mode
to the second aid that promotes humoral immunity by
Single radial immunodiffusion using cooked dishes from EASY
producing dynamics of cellular such as IL-6 and IL-4 and IL
RED company.
cells - 10 B which in turn encourages the production of
• estimate the level of TNFα in a way adsorption linked
immune cells (Gharogozalla et al., 2004; Guo et al., 2003).
immunosorbent Elabscience
MATERIAL & METHODS :
• estimate the kinetic cellular level (IL-6) 6 way linked
Sample collection
immunosorbent adsorption of BOSTER company
Were collected 64 Leather tinge of burn patients admitted
RESULTS AND DISCUSSION:
to the Teaching Hospital Hussein in Karbala province for the
1.
period Mn16-2-2014lgaah 16-2014-8 ages and from
Pseudomonas aeruginosa
different samples were collected by the scanners sterile
Planting Wipes group of patients with burns and environment
disposable which after only two hours planted on different
on media mackcouncy agar and nutritious agar for the initial
agricultural circles (general and selective ) for the purpose of
isolation of the bacteria pseudomonas aeruginosa and after
isolating the bacteria Pseudomonas aeruginosa. Has also
the end of the lap amounting to 24 hours at temperature 37
been collecting blood samples from all patients under study
º m examined developing colonies, which were pale at the
by syringes medical sterile then transfer 2 ml of blood in
center of mackconcy agar a because it is fermented sugar
EDTA containing tubes while leaving 4 ml of blood in plastic
lactose, while the colonies on the central Nutrient large agar
tubes to clot, and blood expel central to separate serum,
her appearance is high and the edges of the flat as well as
which saves in the pipeline epnidroff under freeze 18. Until
similar smell the smell of the grape (Grape like odor) and
an immunological tests of each of the CRP, C3, IL-6 and TNF
color greenish the ability to produce pigment Pyocyanin, then
Isolation and diagnose
quoted the developing germ to the center of strmaid agar
the
bacteria
Pseudomonas
Isolation
and
diagnose
the
bacteria
aeruginosa
for being the center of selectively to those bacteria that
The samples were planting on the media nutritioni agar,
contend Certrimide that inhibit inhibition other bacteria but
blood agar and macounky agar then dishes were incubated
pseudomonas
degree of 37°C for 24 hours then I studied the
appeared cleard on the surface that the middle indication
characteristics of the developing colonies on the circles
as pseudomonas aeruginosa bacterium,
AGRO recipes bacterial cells under an optical microscope
All isolates were holding her many biochemical tests for the
compound after dab with Cram dye and observe the shape,
bacteria developing biochemical tests results as compatible
arrangement and color that studied the biochemical primary
with approved diagnostic systems (Collee, 1996). And after
aeruginosa,
after
incbation
colonies
qualities of the isolates Proceeds for characterization
©SRDE Group, All Rights Reserved.
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Alkateeb D. M. et al., June - July, 2016, 5(4), 2219-2225
the confirmation of the results was the adoption on both
Results also showed a clear increase in the number of blood
API20 and Vetix system.
cells neutrophil as shown in Table (1), where the significant
The results Showed after completion of initial and
increase at the level of probability (p <0.05) for all grades
confirmatory that tests 35 isolation (45%) were the bacteria
simple Incineration (7.35) and average (9.25) and severe
pseudomonas aeruginosa from swabs from patients infected
(11.09) and the control group (3:41). This means that there is
burns The findings come with matching results Alghanimi
an increase in the macrophagic activity in neutrophils in
(2014), which showed that the highest percentage of isolated
patients two to three times more bad eggs, which indicates to
bacterial in burn patients had the bacteria pseudomonas
the evolution of the case of acute burn patients (Paraslow et
aeruginosa , knowing that the thing that increases the chance
al., 2001). As lymphocytes has recorded a significant
of injury that MRSA is a high ability to stay in disinfectants
decline, especially in cases of burns severe, reaching
and liquid medicines or treatments, such as eye drops, as
attribute to 1.379 when compared with the control group
well as anesthesia masks and floor galleries and other
(3.02). The results show the presence of high non-moral in the
processes Ryan and Ray, 2004; Qarah, 2004)). Also, stay in
preparation of the only cell nucleus either all of acidophilus
the hospital for a long time may lead to a high percentage
cells and grassroots there were not significant changes where
of the presence of these bacteria and the increasing risk of
it is believed that the reason for this is due to the fact that
the especially in patients with severe burns Pollack, 2000;
cells stimulate or increase when there is parasitic injury and
Tielker et al., 2005; Japoni et al., 2009)).
hypersensitivity only sensitive and not related to bacterial
2.
injury in General (Doan etal., 2008; Mehta and Hoffbrand,
Complet and differential count for white blood
cells in patients with burns
2009).
The total number of blood cells, increased in burn patients
3.
as compared to control as shown in Figure (1), where he was
Results also showed a clear increase in the number of blood
high for each of the group Statistics burns and moderate to
cells neutrophil as shown in Table (1), where the significant
severe for up to ((10.44, 13.222 and mm3) 15.955 / 103)),
increase at the level of probability (p <0.05) for all grades
respectively, compared to the range control the amount
simple Incineration (7.35) and average (9.25) and severe
(7.87 mm3 / 103) also increase the number of white blood
(11.09) and the control group (3:41). This means that there is
cells fit with the severity of the burn and the size of the
an increase in the macrophagic activity in neutrophils in
psychological pressure on the person burned as the
patients two to three times more bad eggs, which indicates to
psychological effects associated with the burning of a large
the evolution of the case of acute burn patients (Paraslow et
variation in the effect is clear in the immunological criteria
al., 2001). As lymphocytes has recorded a significant
(Demling, 2004)
decline, especially in cases of severe burns,
Figure 1: Shows the total count of white blood cells Leukocyte
in patients and the control group proportion to the degree of
the burn. [L.S.D(0.05)= 2.33]
Figure 2: Complement C3 concentration in healthy subjects
and patients with burns proportion to the degree of the
burn. [L.S.D(0.05)= 12.928]
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C3 complement protein level in burn patients
Int. J. Res. Dev. Pharm. L. Sci.
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Alkateeb D. M. et al., June - July, 2016, 5(4), 2219-2225
Table 1: Differential count of white blood cells in healthy individuals and patients with burn ratios to the point of burning.
Sever burn
M±N
11.095±2.231
1.379±0.526
1.472±0.395
0.067±0.066
0.12±0.043
Patients
Burn degree
Intermediate burn
M±N
9.255±3.125
2.077±0.953
0.947±0.344
0.075±0.038
0.144±0.074
Simple burn
M±N
7.353±2.477
2.487±0.804
0.823±0.451
0.081±0.063
0.17±0.086
control
M±N
Types of WBC cell
3.41 ±1.4
3.02±0.24
0.621±0.17
0.199±0.12
0.11±0.01
Neutrophils
Lymphocytes
Monocytes
Eosinophils
Basophils
reaching attribute to 1.379 when compared with the control
Results of our study are consistent to the findings of a
group (3.02). The results show the presence of high non-
beautiful mechanism (2014) and (kingesely & jones., 2008)
moral in the preparation of the only cell nucleus either all of
who indicated that there is a clear rise in the levels of acute
acidophilus cells and grassroots there were not significant
phase proteins in patients with burns.
changes where it is believed that the reason for this is due to
5.
the fact that cells stimulate or increase when there is parasitic
The results showed of the current study for a rise in cell
injury and hypersensitivity only sensitive and not related to
kinetic values 6 (IL-6) in the sera infected burns compared
bacterial injury in General (Doan etal., 2008; Mehta and
Bash. Also revealed the results of the statistical analysis of
Hoffbrand, 2009).
the existence of significant differences at the level of
4.
probability ( p <0.05) in IL-6 from minor burns and patients
The acute phase C-reactive protein
Cell kinetic level 6 ((IL-6 in patients with burns:
Concentration in burn patients increased acute phase protein
with moderate to severe 118,86,33) pg / ml values),
values in the sera burn patients compared with healthy
respectively, and the control group pg / ml (15) as shown in
people showed in figure ( 3), has revealed the results of
the table and (Figure 4). Current results agreed with Jubouri
statistical analysis of the existence of significant differences
(2008), which confirmed the presence of a rise in the level of
for each degree burn (simple and moderate to severe burn)
cellular motor 6 in patients with burns.
with healthy people at the level of probability( P <0.05).
Some empirical studies have indicated that the tumor factor
The reason for increasing the concentration of the acute
necrotic and IL-1 both types of alpha, beta and IL-6 and IL-
phase protein to increase the concentration of plasma
10 and IL-8 is put up in large quantities in cases of severe
proteins, which increases its focus in cases of inflammation
burns inflamed and the level of asking those cytokines vary
and tissue damage. He has indicated a study by Jeschke and
depending on the type that burns and wounds and the size
others (2013) that the high concentration of CRP is linked
of those burns, as well as the patient's age( Kim etal., 2012).
with the type, size and the severity of those burns.
Infection is an inflammatory response acute with dysfunction
is a complex process of cellular events to control the infection
(Cheung et al., 2008) Kmaan role of developing germ
stimulate macrophage cells mediated by microbial signals
help her launch of inflammatory mediators such as IL-I and IL12 and TNf- and INF- and IL-8, which induce localized
inflammation then will increase blood flow topically to attract
neutrophil cells to the area to kill germs (Munford and Pugin,
2001)
Also, most of the cells in the skin can produce cellular
Figure 3: Acute phase protein concentration (CRP) in healthy
subjects and patients with burns proportion to the degree of
the burn. [L.S.D. (0.05)= 15.81]
©SRDE Group, All Rights Reserved.
dynamics such as stratum and T , macrophages and
fibroblasts, So the impact topically and then travel to the
Hungarian bloody ((Patricia et al., 1999 and this was
Int. J. Res. Dev. Pharm. L. Sci.
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Alkateeb D. M. et al., June - July, 2016, 5(4), 2219-2225
confirmed by researcher Ross (2002) when injected mice
6.
laboratory under the skin with fatty polysaccharide isolated
The results of the current study showed increase the values of
level of kinetic cellular -TNF in burn patients:
from negative bacteria gram and which led to the evolution
kinetic cellular -TNF in sera infected burns compared with
of the case of inflammation the appearance of large
healthy people shown in the figure(5) and) recorded the
amounts of IL-6 in the bloodstream thing that could be due to
highest increase in his patients with severe burns reaching(
the introduction of these quantities produced topically to the
600.45 pg / ml) compared to people healthy (175.15 pg /
Hungarian General the injury severe burns lead to stimulate
ml) as statistical analysis of the existence of significant
the inflammatory response Preliminary to increase production
differences at the level of probability of detection (p <0.05)
and liberation of cytokines inflammatory initial of each of the
in TNF between healthy and groups of patients who suffer
TNF-α IL-1β, and IL-6, which is working on the alert to
from simple burns and moderate to severe values.
prevent the entry of bacterial action pathogen so can be
When you get bruises on the skin bruising
seen rising these factors after 12) -24) hours of injury,
macrophage cells and cells of Langerhans provide antigen
trauma or burn (Gauglitz etal. , 2008; Maass etal., 2002).
(Ag-presentation) to T Tcell cells which works to motivate and
example, the
then start doubling quickly under the influence of the type of
cytokines is IL-1 stimulate cellular dynamics by inflammation,
such as IL - 6, IL - 8, IL-2 and IL-4 and IL-5, which works to
stimulate and doubled the B cells produce antibodies
antibody also launches keratinocytes cells, macrophages also
factor nostril of tumors (TNF-), which contribute to the lifting
of the inflammatory response and the expression of complex
compatibility Histological (MHC) and thus helps to antigen
presenting cells inflammatory (Garcia et al., 2005) in
addition to the fact that the type IL-1 is a factor chemical
attraction (chemotaxis) Direct-white cells, it is in collaboration
with TNF- search expression of adhesion molecules between
Figure 4: The level of cellular motor 6 (IL-6) in patients ratios
to the point of burning. [L.S.D. (0.05) = 11.804]
cells (ICAM ) Intercellular adhesion molecule on epithelial
cells and fibroblasts t that cause adhesion of the white cells
with epithelial cell surfaces (Schumann et al., 1998;
Dinarello, 1997).
Some studies have also indicated that the level of some
interleukin be linked with the size of those burns whenever
there was a large-sized high levels of cytokines and this
speech not for all interleukin. In a study conducted at
Children's Hospital in the state of Texas that the level of
interleukins , INF IL-4, IL-1B, IL-2 was not affected by the
concentrations with increasing size burn area, while the
interleukin other IL-6 level, TNF and IL-8 recorded high
concentrations compared with healthy controls in the case of
children suffering from severe burns (Jeschke etal., 2007).
Some empirical studies have indicated that the tumor factor
Figure 5: Shows the level (TNF) in patients ratios to the point of
necrotic and IL-1 both types of alpha, beta and IL-6 and IL-
burning. [L.S.D. (0.05) = 50.663]
10 and IL-8 is put up in large quantities in cases of severe
burns inflamed and the level of asking those cytokines vary
©SRDE Group, All Rights Reserved.
Int. J. Res. Dev. Pharm. L. Sci.
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Alkateeb D. M. et al., June - July, 2016, 5(4), 2219-2225
depending on the type that burns and wounds and the size
8.
of those burns, as well as the patient's age Kim etal., 2012)).
Also, the role of bacterial infection in cells stimulate
macrophage-mediated microbial signals help her launch of
inflammatory mediators such as IL-I and IL-12 and TNF- and
INF- and IL-8, which induce localized inflammation then the
9.
10.
blood flow will increase topically to attract neutrophil cells to
the area to kill germs (Munford and Pugin, 2001)
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